title: Can bluetongue virus (BTV) be transmitted via caprine embryo transfer?
abstract: The three objectives of this study were to investigate whether cells of early goat embryos isolated from in vivo fertilized goats interact with bluetongue virus (BTV) in vitro, whether the embryonic zona pellucida (ZP) protects early embryo cells from BTV infection, and whether the 10 wash cycles recommended by the International Embryo Transfer Society (IETS) for bovine embryos effectively decontaminates caprine embryos exposed to Bluetongue Virus (BTV) in vitro. Donor goats and bucks were individually screened and tested negative for the virus by RT-PCR detection of BTV RNA in circulating erythrocytes. ZP-free and ZP-intact 8-16 cell embryos were co-cultured for 36 h in an insert over a Vero cell monolayer infected with BTV. Embryos were washed 10 times in accordance with IETS recommendations for ruminant and porcine embryos, before being transferred to an insert on BTV indicator Vero cells for 6 h, to detect any cytopathic effects (CPE). They were then washed and cultured in B2 Ménézo for 24 h. Non-inoculated ZP-free and ZP-intact embryos were submitted to similar treatments and used as controls. The Vero cell monolayer used as feeder cells for BTV inoculated ZP-free and ZP-intact embryos showed cytopathic effects (CPE). BTV was found by RT-qPCR in the ten washes of exposed ZP-free and ZP-intact embryos. In the acellular medium, the early embryonic cells produced at least 10(2.5) TCID(50)/ml. BTV RNA was detected in ZP-free and ZP-intact embryos using RT-qPCR. All of these results clearly demonstrate that caprine early embryonic cells are susceptible to infection with BTV and that infection with this virus is productive. The washing procedure failed to remove BTV, which indicates that BTV could bind to the zona pellucida.
authors.affiliation: LUNAM University, Oniris, (Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering), Department of Research into the Health Risk and Biotechnology of Reproduction UPSP 5301 DGER, France.
authors.forename: M Z Ali
authors.initials: MZ
authors.lastname: Al Ahmad
authors.forename: J L
authors.initials: JL
authors.lastname: Pellerin
authors.forename: M
authors.initials: M
authors.lastname: Larrat
authors.forename: G
authors.initials: G
authors.lastname: Chatagnon
authors.forename: R
authors.initials: R
authors.lastname: Cécile
authors.forename: C
authors.initials: C
authors.lastname: Sailleau
authors.forename: S
authors.initials: S
authors.lastname: Zientara
authors.forename: F
authors.initials: F
authors.lastname: Fieni
mesh_terms: D000818:Animals; D001819:Bluetongue / Q000635:transmission*; D001820:Bluetongue virus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity*; D002522:Chlorocebus aethiops; D046149:Embryo Culture Techniques / Q000662:veterinary; D004624:Embryo Transfer / Q000379:methods / Q000662:veterinary*; D004622:Embryo, Mammalian / Q000821:virology*; D005260:Female; D015511:Goat Diseases / Q000635:transmission* / Q000821:virology; D006041:Goats; D008297:Male; D012367:RNA, Viral / Q000097:blood; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014709:Vero Cells; D015044:Zona Pellucida / Q000821:virology
chemical_list: D012367:RNA, Viral